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Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Integrin beta 1 aa 650-750. The exact sequence is proprietary.
Database link: P05556
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.
Our Abpromise guarantee covers the use of ab52971 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/100000. Detects a band of approximately 140-150 kDa (predicted molecular weight: 88 kDa).
For unpurified, use 1/500.
|IHC-Fr||Use at an assay dependent concentration.|
|IHC-P||1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).
Use of an HRP/AP polymerized secondary antibody is recommended.
For unpurified, use 1/250 - 1/500.
Lanes 1, 5 and 9: Wild-type HAP1 cell lysate (20 µg)
Lanes 2, 6 and 10: Integrin beta 1 knockout HAP1 cell lysate (20 µg)
Lanes 3, 7 and 11: U87-MG cell lysate (20 µg)
Lanes 4, 8 and 12: A431 cell lysate (20 µg)
Lanes 1, 2, 3 and 4: Green signal from target – ab52971 observed at 140 kDa
Lanes 5, 6, 7 and 8: Red signal from loading control – ab8245 observed at 37 kDa
Lanes 9, 10, 11 and 12: Merged (red and green) signal
ab52971 was shown to specifically react with Integrin beta 1 when Integrin beta 1 knockout samples were used. Wild-type and Integrin beta 1 knockout samples were subjected to SDS-PAGE. ab52971 and ab8245 (loading control to GAPDH) were diluted 1/10 000 and 1/2000 respectively and incubated overnight at 4ºC. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
ab52971 at 1/500 staining Integrin beta 1 antibody in human transitional cell carcinoma of bladder by immunohistochemistry (FFPE).
Immunohistochemical analysis of paraffin-embedded human transitional cell carcinoma of bladder tissue labeling Integrin beta 1 with ab52971 at 1/500 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). Counter stained with hematoxylin.
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Integrin beta 1 knockout HAP1 cell lysate (20 µg)
Lane 3: U87-MG cell lysate (20 µg)
Lane 4: A431 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab52971 observed at 140 kDa. Red signal from loading control – ab8245 observed at 37 kDa.
This western blot image is a comparison between ab52971 and a competitor's top cited rabbit polyclonal antibody.
5% NFDM/TBST dilution buffer
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab52971 overnight at 4°C. Antibody binding was detected using ab175781 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
Secondary antibody - anti-rabbit Alexa Fluor 790
Unpurified ab52971 staining human breast cancer metastasis tissue sections by IHC-P. Sections were formaldehyde fixed and subjected to heat mediated antigen retrieval in citrate buffer (pH 6) prior to blocking with a commercial blocking reagent and incubation with the antibody (diluted 1/100) for 18 hours at 4°C. A HRP-conjugated goat anti-rabbit was used as the secondary antibody. This image shows a cancer metastasis at 40x with beta1 staining (in red) in both blood vessels and tumour cells. Blue is Hoechst for nuclei. The antibody detection was enhanced using a commercial Cy3 tyramide signal amplification kit.
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